US6372426B1 - Immunoassay for determining the avidity of immunoglobulins - Google Patents

Immunoassay for determining the avidity of immunoglobulins Download PDF

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Publication number
US6372426B1
US6372426B1 US09/625,059 US62505900A US6372426B1 US 6372426 B1 US6372426 B1 US 6372426B1 US 62505900 A US62505900 A US 62505900A US 6372426 B1 US6372426 B1 US 6372426B1
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Prior art keywords
antibodies
urea
infection
avidity
hydrogen peroxide
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US09/625,059
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Wolfgang Zens
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Siemens Healthcare Diagnostics Products GmbH
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Dade Behring Marburg GmbH
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6854—Immunoglobulins
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/962—Prevention or removal of interfering materials or reactants or other treatment to enhance results, e.g. determining or preventing nonspecific binding

Definitions

  • the present invention relates to a method and a diagnostic aid for the qualitative or quantitative detection of antibodies and for determining their avidity. This makes it possible to diagnose the early phase of viral, bacterial or parasitic infections.
  • the diagnostic aid according to the invention is particularly suitable for automated processing in large analytical laboratories.
  • Immunoassays are frequently employed, because of their particularly good specificity and sensitivity, for detecting immunoglobulins in serum and plasma samples for medical diagnostic purposes. In addition, immunoassays are distinguished by being simple to use.
  • urea The substance which is mostly used, urea, is, at the concentration recommended by the authors, on the point of crystallization, which results in frequent blockage of all pipette tips and tubes of equipment for automatic or partly automatic processing of immunoassays (immunoassay processors).
  • the present invention therefore relates to a method for the qualitative or quantitative detection of an antibody, in which this antibody is brought into contact with the antigen against which it is directed so that immune complexes are able to form, and in which the reaction mixture is brought into contact with a protein-denaturing agent which destabilizes immune complexes containing antibodies of low avidity, while immune complexes containing antibodies of higher avidity are substantially retained, and in which the extent of the binding of the antibody to the antigen is determined by a method known to the skilled worker, wherein the protein-denaturing agent is urea-hydrogen peroxide.
  • the present invention additionally relates to a method for determining the avidity of an antibody, in which the antibody is brought into contact in a first and a second mixture independently of one another with the antigen against which the antibody is directed so that immune complexes are able to form, and in which one of the two mixtures is brought into contact with a protein-denaturing agent which destabilizes immune complexes containing antibodies of low avidity, while immune complexes containing antibodies of higher avidity are substantially retained, and in which the extent of the binding of the antibody to the antigen in both samples is determined independently of one another by a method known to the skilled worker, and where the avidity of the antibody is revealed by the ratio of the extent of the antigen-antibody bindings in the first and the second mixture, wherein the protein-denaturing agent is urea-hydrogen peroxide.
  • a preferred method of this type is one in which the antigen is brought into contact, in a form bound to a solid phase, with the antibody, and subsequently washed with a buffer solution containing urea-hydrogen peroxide.
  • preferred methods of this type include the enzyme immunoassay, radioimmunoassay, Western blot or immunofluorescence assay.
  • the skilled worker is aware of other immunoassay systems which can be carried out straightforwardly according to the invention using urea-hydrogen peroxide by means of the present is description.
  • Antibodies of this type may be directed against viruses, bacteria or parasites such as, for example, EBV, rubella virus, CMV, hantavirus, parvovirus B19, VZV, HHV 6, HBV, HCV, HIV, RSV, HSV-1, HSV-2 or Toxoplasma gondii.
  • the determination of avidity can advantageously be carried out using commercially obtainable immunoassays (for example ELISA), partly or fully automatically.
  • Another advantage of the present invention is the very good interpretability of the results.
  • the novel method for determining the avidity of antibodies is particularly suitable for differentiating fresh (i.e. only recently occurring) infections from older (i.e. less recent) infections.
  • the present invention relates to methods for detecting an acute rubella virus infection, or one which has recently completed its course, a first CMV infection, first EBV infection, first HSV infection or Toxoplasma gondii infection.
  • the present invention additionally relates to diagnostic aids (diagnostic reagents, assay kits) which are suitable for application of the novel methods described. Diagnostic aids of these types are produced in a manner known per se to the skilled worker, based on the present description of the invention.
  • FIG. 1 shows molarity optimization tests with the protein-denaturing substances urea and urea-hydrogen peroxide in the Enzygnost R anti-rubella virus IgG ELISA.
  • Top panel serum Ab05, serum taken 4 weeks after rubella infection.
  • Bottom panel serum 281, late phase, several years after contact with rubella virus.
  • the wash solution from commercially obtainable ELISA kits is frequently used as diluent solution for protein-denaturing substances.
  • POD wash solution Order No. OSEW, Behring Diagnostics GmbH, Marburg, Germany.
  • any other wash solution suitable in principle for washing an ELISA could have been used.
  • the urea-hydrogen peroxide is used in the range between 2.5 mol/l and 6.5 mol/l, preferably between 4.5 and 6.0 mol/l.
  • the optimized concentration is 5.3 mol/l. It is also conceivable to use urea-hydrogen peroxide in other solutions used for carrying out immunoassays, for example in sample dilution buffers.
  • novel urea-hydrogen peroxide solution is used to carry out an immunoassay, for example an ELISA, by a conventional method.
  • the modification of the novel method by comparison with known methods is that incubation of the sample is followed by washing twice with a volume suited to the method (for example about 0.3 ml in an ELISA based on microtitre plates) of wash solution containing urea-hydrogen peroxide, and subsequently, for example, washing twice with in each case the same volume of wash solution.
  • a volume suited to the method for example about 0.3 ml in an ELISA based on microtitre plates
  • wash solution containing urea-hydrogen peroxide containing urea-hydrogen peroxide
  • the assay is carried out, if automatic operation is required, on an ELISA processor (obtainable, for example, from Behring Diagnostics GmbH).
  • FIG. 1 shows that the novel urea-hydrogen peroxide solution in a concentration range from 2.5 to 6.5 mol/l achieves the best separation between the two groups of sera.
  • the discrimination of the method is to be shown in a group of confirmed rubella IgM-positive sera comparing with a group of confirmed rubella IgM-negative sera.
  • HSV herpes simplex virus
  • herpes simplex sera from the late phase of infection have an average avidity index of greater than about 50%.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Pathology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • General Physics & Mathematics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Peptides Or Proteins (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
US09/625,059 1997-05-02 2000-07-24 Immunoassay for determining the avidity of immunoglobulins Expired - Fee Related US6372426B1 (en)

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US09/625,059 US6372426B1 (en) 1997-05-02 2000-07-24 Immunoassay for determining the avidity of immunoglobulins

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
DE19718361 1997-05-02
DE19718361A DE19718361A1 (de) 1997-05-02 1997-05-02 Immunoassay zur Aviditätsbestimmung von Immunglobulinen
US7129298A 1998-05-01 1998-05-01
US09/625,059 US6372426B1 (en) 1997-05-02 2000-07-24 Immunoassay for determining the avidity of immunoglobulins

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US7129298A Division 1997-05-02 1998-05-01

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US (1) US6372426B1 (de)
EP (1) EP0875761B1 (de)
JP (1) JP3917753B2 (de)
AT (1) ATE199783T1 (de)
CA (1) CA2236554C (de)
DE (2) DE19718361A1 (de)
ES (1) ES2155709T3 (de)

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060194195A1 (en) * 2003-04-11 2006-08-31 Kalish Marcia L Multiple antigenic peptide assay for detection of hiv or siv type retroviruses
US20070099295A1 (en) * 2005-11-02 2007-05-03 Maine Gregory T Methods for the determination of antibody IgG avidity
WO2012170765A2 (en) 2011-06-10 2012-12-13 Oregon Health & Science University Cmv glycoproteins and recombinant vectors
EP2568289A2 (de) 2011-09-12 2013-03-13 International AIDS Vaccine Initiative Immunselektion von rekombinantem vesikulärem Stomatitisvirus mit Expression von HIV-1-Proteinen durch Breitbandneutralisierungs-Antikörper
EP2586461A1 (de) 2011-10-27 2013-05-01 Christopher L. Parks Von einem eingehüllten Virus abgeleitete Virenpartikel
EP2679596A1 (de) 2012-06-27 2014-01-01 Simon Hoffenberg HIV-1 Env-Proteinvariante
CN104330553A (zh) * 2014-11-20 2015-02-04 扬州大学 一种无标记化学发光免疫传感器及其免疫分析方法
EP2848937A1 (de) 2013-09-05 2015-03-18 International Aids Vaccine Initiative Verfahren zur Identifizierung neuartiger HIV-1-Immunogene
EP2873423A2 (de) 2013-10-07 2015-05-20 International Aids Vaccine Initiative Lösliche hiv-1-hüllglykoproteintrimere
EP3069730A2 (de) 2015-03-20 2016-09-21 International Aids Vaccine Initiative Lösliche hiv-1-hüllglykoproteintrimere
EP3072901A1 (de) 2015-03-23 2016-09-28 International Aids Vaccine Initiative Lösliche hiv-1-hüllglykoproteintrimere
EP3187585A1 (de) 2010-03-25 2017-07-05 Oregon Health&Science University Cmv-glycoproteine und rekombinante vektoren

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE19910045A1 (de) * 1999-03-08 2000-09-14 Mikrogen Molekularbiol Entw Verfahren zur Bestimmung der Avidität von Antikörpern
EP1237928A1 (de) * 1999-12-13 2002-09-11 ZLB Bioplasma AG Verbesserung des therapeutischen potential von immunoglobulinpreparationen
DE10232203A1 (de) * 2002-07-16 2004-02-19 Institut Virion/Serion Gmbh Verfahren und Diagnostikum zum qualitativen oder quantitativen Nachweis von Antikörpern und zur Bestimmung von deren Avidität
EP3961215A1 (de) * 2020-08-25 2022-03-02 Mikrogen GmbH Verfahren zur bestimmung der avidität von gegen coronavirus gerichteten antikörpern sowie hierzu geeignete testkits

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS62220865A (ja) 1986-03-24 1987-09-29 Yatoron:Kk 均一系酵素免疫学的測定方法
WO1990002202A1 (en) 1988-08-16 1990-03-08 Cetus Corporation Reduction of peroxidatic and catalatic interference with assays of peroxidatic activity
US5183901A (en) 1992-01-24 1993-02-02 Isp Investments Inc. Urea-hydrogen peroxide-polyvinylpyrrolidone
US5206385A (en) 1992-01-24 1993-04-27 Isp Investments Inc. Urea-hydrogen peroxide-polyvinylpyrrolidone process
US5512659A (en) 1989-08-04 1996-04-30 Syntex (U.S.A.) Inc. Compositions useful in heterogeneous immunoassays
WO1997009619A1 (en) 1995-09-01 1997-03-13 Johnson & Johnson Clinical Diagnostics, Inc. Analytical element and method for the determination of a specific binding ligand using a vanadium bromoperoxidase as a signal-generating enzyme
US5679537A (en) 1994-10-26 1997-10-21 Mcgill University Immunoassays for measuring the avidity of rheumatoid factor in rheumatoid arthritis
US5830634A (en) 1994-02-23 1998-11-03 Dade Behring Marburg Gmbh Peptides derived from a retrovirus of the HIV group and their use

Patent Citations (8)

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JPS62220865A (ja) 1986-03-24 1987-09-29 Yatoron:Kk 均一系酵素免疫学的測定方法
WO1990002202A1 (en) 1988-08-16 1990-03-08 Cetus Corporation Reduction of peroxidatic and catalatic interference with assays of peroxidatic activity
US5512659A (en) 1989-08-04 1996-04-30 Syntex (U.S.A.) Inc. Compositions useful in heterogeneous immunoassays
US5183901A (en) 1992-01-24 1993-02-02 Isp Investments Inc. Urea-hydrogen peroxide-polyvinylpyrrolidone
US5206385A (en) 1992-01-24 1993-04-27 Isp Investments Inc. Urea-hydrogen peroxide-polyvinylpyrrolidone process
US5830634A (en) 1994-02-23 1998-11-03 Dade Behring Marburg Gmbh Peptides derived from a retrovirus of the HIV group and their use
US5679537A (en) 1994-10-26 1997-10-21 Mcgill University Immunoassays for measuring the avidity of rheumatoid factor in rheumatoid arthritis
WO1997009619A1 (en) 1995-09-01 1997-03-13 Johnson & Johnson Clinical Diagnostics, Inc. Analytical element and method for the determination of a specific binding ligand using a vanadium bromoperoxidase as a signal-generating enzyme

Non-Patent Citations (6)

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Title
Behring Diagnostics GMBH, Order NO OSEW, Marburg, Germany.
Hillar O. Kangro , et al. "Antibody Avidity Following Varicella-Zoster Virus Infections", Journal of Medical Virology, vol. 33, pp. 100-105, 1991.
J. Polance, et al., "Evaluation of Protein-Denaturing Immunoassays for Avidity of Immunoglobulin G to Rubella Virus", Journal of Clinical Laboratory Analysis, vol. 8, pp. 16-21, 1994.
J. Schubert, et al., "Avidity Determination in Epstein-Barr Virus Diagnosis-a Comparison of Immunofluorescence Assay and ELISA", J. Lab Med., vol. 20, No. 12, pp. 713-717, 1996.
J.J. Gray, "Avidityof EBV VCA-specific IgG antibodies: distinction between recent primary infection, past Infection and reactivation", Journal of Virological Methods, vol. 52, pp. 95-104, 1995.
Zusammenfassung, Patent Abstracts of Japan, vol. 012, No. 088 (P-678), Mar. 23, 1988 & JP 62 220865 A (Yatoron :kk), Sep. 29, 1987.

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8524461B2 (en) 2003-04-11 2013-09-03 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Multiple antigenic peptide assay for detection of HIV or SIV type retroviruses
US20100222236A1 (en) * 2003-04-11 2010-09-02 The Government of The United States of America, as Represented by the Secretary, Department Multiple antigenic peptide assay for detection of hiv or siv type retroviruses
US20060194195A1 (en) * 2003-04-11 2006-08-31 Kalish Marcia L Multiple antigenic peptide assay for detection of hiv or siv type retroviruses
WO2007056064A1 (en) * 2005-11-02 2007-05-18 Abbott Laboratories Methods for the determination of antibody igg avidity
US7432046B2 (en) 2005-11-02 2008-10-07 Abbott Laboratories Methods for the determination of antibody IgG avidity
US20070099295A1 (en) * 2005-11-02 2007-05-03 Maine Gregory T Methods for the determination of antibody IgG avidity
EP3187585A1 (de) 2010-03-25 2017-07-05 Oregon Health&Science University Cmv-glycoproteine und rekombinante vektoren
WO2012170765A2 (en) 2011-06-10 2012-12-13 Oregon Health & Science University Cmv glycoproteins and recombinant vectors
EP2568289A2 (de) 2011-09-12 2013-03-13 International AIDS Vaccine Initiative Immunselektion von rekombinantem vesikulärem Stomatitisvirus mit Expression von HIV-1-Proteinen durch Breitbandneutralisierungs-Antikörper
EP2586461A1 (de) 2011-10-27 2013-05-01 Christopher L. Parks Von einem eingehüllten Virus abgeleitete Virenpartikel
EP2679596A1 (de) 2012-06-27 2014-01-01 Simon Hoffenberg HIV-1 Env-Proteinvariante
EP2848937A1 (de) 2013-09-05 2015-03-18 International Aids Vaccine Initiative Verfahren zur Identifizierung neuartiger HIV-1-Immunogene
EP2873423A2 (de) 2013-10-07 2015-05-20 International Aids Vaccine Initiative Lösliche hiv-1-hüllglykoproteintrimere
CN104330553A (zh) * 2014-11-20 2015-02-04 扬州大学 一种无标记化学发光免疫传感器及其免疫分析方法
EP3069730A2 (de) 2015-03-20 2016-09-21 International Aids Vaccine Initiative Lösliche hiv-1-hüllglykoproteintrimere
EP3072901A1 (de) 2015-03-23 2016-09-28 International Aids Vaccine Initiative Lösliche hiv-1-hüllglykoproteintrimere

Also Published As

Publication number Publication date
DE59800519D1 (de) 2001-04-19
JPH10319016A (ja) 1998-12-04
CA2236554A1 (en) 1998-11-02
EP0875761B1 (de) 2001-03-14
CA2236554C (en) 2008-12-02
ES2155709T3 (es) 2001-05-16
ATE199783T1 (de) 2001-03-15
EP0875761A1 (de) 1998-11-04
DE19718361A1 (de) 1998-11-05
JP3917753B2 (ja) 2007-05-23

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